A single dominant gene for downy mildew resistance in broccoli

Downy mildew, incited by Peronospora parasitica (Pers.: Fr.) Fr., is a destructive disease of broccoli (Brassica oleraceaL., Italica Group). Resistant cultivars represent a desirable control method to provide a practical, environmentally benign, and long-term means of limiting damage from this disease. Doubled-haploid (DH) lines developed by us exhibit a high level of downy mildew resistance at the cotyledon stage. To determine the mode of inheritance for this resistance, a resistant DH line was crossed to a susceptible DH line to make an F₁, from which F₂ and backcross (BC) populations were developed. All populations were evaluated for response to artificial inoculation with P. parasitica at the cotyledon stage. All F₁ plants (including reciprocals) were as resistant as the resistant parent, indicating no maternal effect for this trait. F₂ populations segregated approximately 3resistant to 1 susceptible, BC populations using the resistant parent as the recurrent parent contained all resistant plants, and the BC to the susceptible parent segregated 1 resistant to 1 susceptible. These results indicate that resistance is controlled by a single dominant gene. This gene should be easily incorporated into F₁ hybrids and used commercially to prevent downy mildew at the cotyledon stage. This revised version was published online in August 2006 with corrections to the Cover Date.     

Inheritance of resistance to black rot (Xanthomonas campestris pv. Campestris) in cauliflower (Brassica oleracea var. Botrytis)

Summary Black rot disease caused by Xanthomonas campestris pv. campestris is a limiting factor in the commercial production of the cauliflower crop. Crosses were attempted between SN 445, a mid season cultivar resistant to black rot and two highly susceptible commercial cultivars (Pusa Snowball-1 and K-1). Studies of the F₁'s, F₂'s and back crosses indicated that SN 445, carries a dominant gene imparting resistance to black rot.     

Differential response and genes for resistance to Peronospora parasitica (downy mildew) in Brassica juncea (mustard)

The response of a wide range of Brassica juncea accessions to 14 isolates of Peronospora parasitica, 12 from India (IP00A, IP02, IP03, IP04, IP04A, IP05, IP05B, IP33 and IP33A were derived from B. juncea; IP09, IP14 and IP13A from B. rapa) and two from B. napus in the UK (R1 and P003), was screened. Sixteen differential host response groups to these isolates (classified as groups A‐P) were identified. Groups‘A’and‘B’expressed the widest resistance profiles to these isolates. Group‘A’was susceptible to isolates IP05 and IP05B, moderately resistant to isolate IP33 and resistant to all other isolates. Group‘B’was susceptible to isolates IP03, IP04 and IP04A, and resistant to the other isolates. Putative homozygous lines resistant to all 14 isolates were selected from the F₄ progeny of crosses involving lines RESBJ‐200 from group‘A’(selection from cv. Kranti) and RESBJ‐190 from group‘B’(selection from cv. Krishna). Both selections were selfed and tested for uniformity of reactions to all isolates for three generations. The resistance of RESBJ‐200 to isolates IP00A, IP04A and IP33A seems to be conditioned by single dominant genes. The resistance of RESBJ‐190 to isolates IP00A, IP05B and IP33A was also conditioned by single dominant genes. The gene for resistance to IP00A and IP33A in RESBJ‐200 seems to be independent of the genes for resistance to the same isolates in RESBJ‐190. The new genes for differential resistance to P. parasitica will be of value in future studies of the genetics of the host‐pathogen interaction and for breeding for disease resistance.     

RAPD markers linked to resistance to downy mildew disease in soybean

To determine and utilize RAPD markers linked to resistance to downymildew incited by Peronospora manshurica in soybean, a resistantcultivar `AGS129' was crossed to a susceptible cultivar `Nakhon Sawan 1'(NS1). F₂ and BC₁ populations were advanced from the F₁ and evaluatedfor resistance to the disease. ²-test demonstrated that the resistancewas controlled by a single dominant gene (Rpmx). Near-isogenic lines(NILs) and bulked segregant analysis (BSA) were used to identify RAPDmarkers linked to the gene. Six DNA bulks namely F₅(R), F₅(S),BC₆F₃(R), BC₆F₃(S), F₂(R) and F₂(S) were set up by pooling equalamount of DNA from 8 randomly selected plants of each disease responsetype. A total of 180 random sequence decamer oligonucleotide primerswere used for RAPD analysis. Primer OPH-02 (5 TCGGACGTGA 3 andOPP-10 (5 TCCCGCCTAC 3) generated OPH-02₁₂₅₀ and OPP-10₈₃₁fragments in donor parent and resistant bulks, but not in the recurrentparent and susceptible ones. Co-segregation analysis using 102 segregatingF₂ progenies confirmed that both markers were linked to the Rpmxgene controlling downy mildew disease resistance with a genetic distance of4.9 cm and 23.1 cm, respectively. Marker OPH-02₁₂₅₀ was presentin 13 of 16 resistant soybean cultivars and absent in susceptible cultivars,thus confirming a potential for MAS outside the mapping population.     

Linkage in Spinacia oleracea L. of two race-specific genes for resistance to downy mildew Peronospora farinosa f. sp. spinaciaeByford

Summary The genetics of resistance to races 1 and 2 of downy mildew Peronospora farinosa f. sp. spinaciae in the spinach variety Nores was investigated in crosses and backcrosses with the susceptible variety Eerste Oogst. Resistance appears to be governed by two closely linked genes in coupling phase with a recombination percentage of 4.6.     

Selection for partial resistance to downy mildew in peas by means of greenhouse tests

Summary A partially downy mildew resistant pea line was back-crossed to a susceptible cultivar with more pods per node and lower seed weight. Breeding lines with different degrees of infection in a greenhouse test were selected. These lines and the two parental lines were investigated in field trials and tested in the greenhouse for four generations. Significant genetic variation among lines was found for infection of seedlings in greenhouse tests, and infection of pods, pod set and seed weight in field-trials. Infection of seedlings in the greenhouse was correlated with infection of pods in the field. In greenhouse tests, the non-genetic variance component was large in comparison with the genetic component and a significant genotype trial interaction was found. Significant repeatability was obtained for downy mildew on seedlings and pods, number of pods per node and seed weight. An unfavourable correlation between susceptibility to downy mildew and number of pods per node was found. No single breeding line showed the ideal combination of good resistance, high number of pods per node and small seeds. However, one line showing better resistance than the susceptible parent, with smaller seeds and more pods per node than the resistant parent was found. The susceptible parent also carries some resistance factor that is not present in the resistant parent.     

The identification and characteristics of field resistance to lettuce downy mildew (Bremia lactucae Regel)

Summary Eighty-one accessions of three Lactuca species which showed no recognisable race specific resistance to Bremia lactucae when tested in the laboratory as seedlings, exhibited different degrees of susceptibility when exposed to natural field infection. As a group, crisp genotypes had less mildew and a slower rate of disease development than other types of lettuce. Wild forms of Lactuca sativa and Lactuca serriola were particularly susceptible. In a further trial, the low field susceptibility of three lettuce cultivars (Iceberg, Batavia blonde de Paris and Grand Rapids) was confirmed. Disease development on cv. Iceberg was compared to that on the highly susceptible cv. Hilde in experiments where the two cultivars were grown either in close proximity or in isolation. The absolute level of attack on cv. Iceberg depended upon disease pressure and differences between the two types only became apparent approximately 8 wk after sowing.     

The role of resistance breeding in the intergrated control of downy mildew (Bremia lactucae) in protected lettuce

Summary Over the last 30 years, six resistance alleles (Dm2, Dm3, Dm6, Dm7, Dm11 andDm16) located in two linkage groups, have contributed to the control of downy mildew in lettuce crops grown under protection (glass or polythene) in northern Europe. More recently, an as yet genetically uncharacterised resistance factor, R18, has also begun to assume importance. The occurrence of the various combinations of these resistance alleles that exist in commercial cultivars has been dictated by the pathotypes ofBremia lactucae used in their selection but also restricted by linkage in repulsion. In the UK, a pathotype ofB. lactucae insensitive to phenylamide fungicides, such as metalaxyl, emerged in 1978 and became prevalent throughout lettuce production areas in subsequent years. The specific virulence of this pathotype was identical to the previously described phenylamide sensitive pathotype NL10 and cultivars carryingDm11, Dm16 or R18 were resistant. Consequently, an integrated control strategy based on the utilisation of metalaxyl on cultivars carryingDm11 provided effective control in UK until 1987 when a new phenylamide insensitive pathotype began to cause problems. The specific virulence of this second pathotype, which was first reported in the Netherlands and France, was identical to the previously described phenylamide sensitive pathotype NL15. Cultivars carryingDm6, Dm16 or R18, but notDm11, were resistant to NL15; consequently an appropriate change in the cultivar recommendations for use in the integrated control strategy was successfully promulgated. It is predicted that variations of this integrated control strategy involving the use of appropriately selectedDm gene combinations may prove effective for some time. This prediction is based on studies of the status of the avirulence loci in the two phenylamide insensitive pathotypes and of the specific virulence characteristics of phenylamide sensitive components of the pathogen population.     

A demonstration of the inheritance of field resistance to lettuce downy mildew (Bremia lactucae Regel.) in progeny derived from cv. Grand Rapids

Summary The inheritance of field resistance to downy mildew in lettuce derived from cv. Grand Rapids was studied in F₂, F₃ and F₄ progeny from a cross with the more susceptible cv. Lobjoits Green Cos. The results for 32 randomly produced F₃ families and the F₂ population corresponded closely although the absolute level of resistance varied from season to season according to disease pressure and environmental conditions. The field resistance character showed a quantitative pattern of inheritance but the rapid response to selection at F₃ implied that only a few genes may be involved. F₄ families raised from families selected from the two extremes of the F₃ distribution fell into distinct categories with resistance levels corresponding to that of their F₃ parents. An association between resistance and morphological features of the resistant parent was present at F₂ but not in subsequent generations when individuals and families which were as resistant as cv. Grand Rapids and yet bore little morphological resemblance to this cultivar were readily recovered.     

Colocation of downy mildew (Plasmopara halstedii) resistance genes in sunflower (Helianthus annuus L.)

Summary The Pl6 locus in the inbred sunflower (Helianthus annuus L.) line HA335 giving resistance to French races of downy mildew (Plasmopara halstedii (Farl.) Berl. & de Toni. was localized by molecular techniques. A bulked segregant analysis was made on the F2 progeny from a cross between this line and H52, a downy mildew susceptible line. The resistance gene in HA335 was found to have the same linked RFLP marker loci as those determined for Pl1 (resistance to race 1 in the line RHA266) on linkage group 1 of the consensus RFLP map of the cultivated sunflower. Pl1 and Pl6 thus appear either to be allelic or closely linked. The implications for sunflower breeding are discussed.     

Expression of resistance to downy mildew at cotyledon and adult plant stages in Brassica oleracea L.

This research compared the expression of resistance to downy mildew at cotyledon and adult plant stages in seven Brassica oleracea genotypes against two P. parasitica isolates of different virulence. Seven day old seedlings were dual inoculated under controlled environment by depositing two 10 μl droplets of a spore suspension of a different isolate on each cotyledon and the interaction phenotype (IP) evaluated 7 days later using a 0–5 scale of increasing susceptibility. The seedlings were transplanted to 16 cm pots and grown in the greenhouse for 110 days (15 to 28 leaves). Adult plants were tested using a single leaf inoculation method that allowed the same plant to be simultaneously inoculated with the two P. parasitica isolates. Leaves were scored 10 days after inoculation using a 0–5 scale of increasing susceptibility. The inoculation of the same plant with the two isolates produced different combinations of cotyledon (CT) and adult-plant (AP) interaction-phenotype according to the genotype. CrGC 3.1 was susceptible at CT and AP stages, ‘Algarvia’ resistant at CT and AP stages, and broccoli ‘A’ susceptible at CT and resistant at AP stage against the two isolates. ‘Murciana’ and broccoli ‘B’ were differential at CT and AP stages. Savoy and Shetland cabbages were differential at CT stage and resistant to both isolates as adult plants. Cotyledon resistance could not be used to predict adult-plant resistance since the two types of resistance were very poorly correlated. Resistance can be race specific either at cotyledon or at adult-plant stage. This revised version was published online in July 2006 with corrections to the Cover Date.     

Evaluation of greenhouse inoculation techniques to screen sorghum for resistance to downy mildew

Summary Six inoculation techniques were compared for the artificial promotion of downy mildew (Peronosclerospora sorghi) in sorghum. These were (1) sprouted seeds incubated between sporulating infected leaves, (2) sprouted seeds depped in conidial suspension, (3) sprouted seeds sprayed with conidial suspension, (4) seedlings at plumule stage inoculated with drops of a conidial suspension, (5) seedlings at plumule stage sprayed with a conidial suspension, and (6) seedling showered with conidia falling from infected leaves. Seedlings at the one-leaf stage sprayed with a conidial suspension (6 × 10⁵ ml^-1) showed the highest systemic infection (100%) in the susceptible lines IS 643 and IS 18433. This technique is effective, repeatable, and allows the deposition of a conidial suspension as a fine mist on the entire seedling surface. In the greenhouse, the technique was used to test the downy mildew reaction of genotypes previously reported as resistant (< 5% incidence) in 3–4 years of field screenings. Of the 61 genotypes tested, 21 were free from downy mildew, 14 had less than 5% incidence, and the rest showed variable susceptible reactions. Therefore, the technique can be reliably and effectively used in the greenhouse to detect disease escapes and to indentify resistance.     

The location and characteristics of novel sources of resistance to Bremia Lactucae Regel (downy mildew) in wild Lactuca L. Species

Summary Twenty-one accessions of 3 wild Lactuca species which could be hybridised with L. sativa, the cultivated lettuce, were inoculated at different stages of plant development with 3 multivirulent isolates of Bremia lactucae. Nineteen sources of resistance to B. lactucae, not attributable to the previously recognised resistance factors 1–11 were identified. Two lines of L. serriola showed similar resistance patterns as lines carrying R11. The resistance of some accession was incomplete particularly at the seedling stage and this phenomenon may be race specific.Tests on segregating F₂ populations of crosses between 2 different L. serriola accessions and L. sativa cultivars showed that the resistance in one line (LSE/18) appears to be inherited as a single dominant gene, which is sometimes incomplete in expression and allelic to either Dm6 or R7. The segregation patterns for resistance in PI 281876 did not give readily interpretable ratios.To assess the frequency of occurrence in B. lactucae populations of virulence factors to overcome this novel resistance, 11 of the novel sources of resistance were inoculated with numerous collections of the pathogen from the UK, Czechoslovakia and elsewhere and found to show a high level of resistance.     

Genetic variability for resistance to sorghum downy mildew (Peronosclerospora sorghi) and Rajasthan downy mildew (P. heteropogoni) in the tropical/sub-tropical Asian maize germplasm

Downy mildews cause considerable damage to maize (Zea mays L.) worldwide, particularly in the tropical Asia. We have evaluated a set of 42 tropical/sub-tropical maize inbred lines developed in different countries in Asia (India, Thailand and Philippines), and Mexico, for analysing the genetic variability for resistance to sorghum downy mildew [Peronosclerospora sorghi; SDM] and Rajasthan downy mildew [P. heteropogoni; RDM]. Experiments were carried out in replicated trials under artificial infection in field conditions against SDM and RDM at Mandya in Karnataka, India, and Udaipur in Rajasthan, India, respectively, during 1999 and 2000. The study resulted in identification of five inbred lines offering consistent and strong resistance to both SDM and RDM, while several inbred lines revealed resistance only to RDM. It was also revealed that the SDM-resistant inbreds are invariably resistant to RDM, while the RDM-resistant inbreds might show differential responses to the SDM. The maize inbred lines identified in this study with broad-spectrum resistance to downy mildews could be potentially useful for basic and applied research work on downy mildews in tropical Asia. This revised version was published online in August 2006 with corrections to the Cover Date.     

An association between resistance to root aphid (Pemphigus bursarius L.) and downy mildew (Bremia lactucae Regel) in lettuce

Summary Many lettuce cultivars (Lactuca sativa L.) with high resistance to lettuce root aphid (Pemphigus bursarius L.) also carried the gene Dm-6 for specific resistance to downy mildew (Bremia lactucae Regel). This suggests the possibility of linkage between this gene and root aphid resistance. The origin of this association is discussed.     

Selection for brown stripe downy mildew resistance in maize

Summary The maize (Zea mays L.) cultivar Makki Safed 1 (MS1) with susceptibility to brown stripe downy mildew (BSDM) caused by Sclerophthora rayssiae var. zeae Payak & Renfro, was subjected to two cycles of mass selection and one cycle of full-sib family selection. Selection was carried out primarily for BSDM resistance.The mass selection was practised under artificial epiphytotic conditions in a disease nursery. Full-sib progenies and performance trials on MS1 and its improved versions were grown in diseased and disease free environments.Mass selection resulted in a significant improvement for resistance to BSDM. A cycle of full-sib selection resulted in an additional improvement for resistance to the disease. The disease rating of the improved version was 1.5 against 4.5 for the original population (scale: 1, highly resistant to 5, highly susceptible). The yield of the improved populations of MS1 was significantly greater than that of MS1 in the disease nursery. In disease free experiments, the improved populations showed almost no yield advantage over MS1. There were also no significant differences between the original population and the improved population after three selection cycles for ear length, ear girth, number of kernel rows per ear, number of kernels per row, 1000-kernel weight, plant height, ear height and days to silk.     

Partial resistance in lettuce to downy mildew (Bremia lactucae). 3. Correspondence between resistance levels of cotyledons and leaf discs and resistance of adult plants

Summary The correlation between resistance levels of seedlings with cotyledons only and leaf discs taken from young and old leaves on the one hand and the resistance of adult plants in bicyclic field tests on the other was investigated. Components of resistance were: latent period (LP) the percentage of seedlings (SEED) or the number of cotyledons (COT) infected, the number of sporangiophores (SPOR) per cotyledon or leaf dise and the % of the surface (SURF) per cotyledon or disc covered with sporangiophores.For both cotyledons and leaf discs, significant differences occurred between plant genotypes for LP, SPOR and SURF. The correspondence between results of cotyledon testing and field testing was generally negligible. There was a rather good correlation between the resistance levels of leaf discs, expressed in SPOR and SURF and the resistance of adult plants. On the basis of a smaller coefficient of variation for SPOR this parameter should be preferred to predict the resistance of adult plants by testing leaf discs.     

Inheritance of resistance to powdery mildew (Erysiphe graminis f. sp. hordei) in eleven primitive barley varieties

The inheritance of the powdery mildew resistance of eleven primitive barley varieties was investigated. crosses with varieties with identified resistance genes revealed that at least three different genes are present in a group of six genotypes from Greece (Hor736, Hor847, Hor878, Hor1159, Hor1379 and Hor1873) and one from Turkey (Hor1188). One dominant gene is common to these seve genotypes and in the Greek accessions a second gene is present. The expression of this second gene was respectively dominant in the crosses with Hor736 and Hor1159, recessive in the crosses with Hor878 and Hor1873, and depended on the employed powdery mildew isolate in the crosses with Hor847 and Hor1379. The detected genes differ from the resistance genes of the crossing partners, viz. Ml-al2, Ml-(Ab), Ml-(CP) and Ml-(1402). The accessions from China (Hor824 and Hor4021) each possess two resistance genes which differ from Ml-a12, Ml-(La) and Ml-(CP). The major gene is common to both accessions and is either identical, allelic or closely linked to a gene in the variety Nigrate. The Columbian accession Hor1894 possesses one resistance gene linked to a resistance gene in Nigrate but this gene differs from the Ml-a locus. The variety Palestine (Hor3997) possesses two resistance genes of which one is allelic or closely linked to Ml-(at).     

Inheritance of stalk rot resistance in cauliflower (Brassica oleracea var. Botrytis L.)

Summary The inheritance of resistance in cauliflower to stalk rot (Sclerotinia sclerotiorum (Lib.) de Bary) was investigated in population from six generations of six crosses. Disease incidence was recorded on 4 parents, 6 F^s ₁, 6 F^s ₂ and 12 back-crosses in a screenhouse under artificially created epiphytotic conditions. Resistance to stalk rot in this set of parents was found to be polygenic and under the control of recessive genes and due primarily to additive gene action. A breeding strategy emphasizing recurrent selection should lead to improvement in resistance.